title: Effect of salts on Azotobacter vinelandii nitrogenase activities. Inhibition of iron chelation and substrate reduction.
abstract: The effect of salts on the catalytic activity of the molybdenum-containing nitrogenase complex from Azotobacter vinelandii has been investigated. NaCl was found to inhibit the reduction of the substrates, protons, acetylene, and dinitrogen by a common mechanism. The pattern of inhibition is sigmoidal, indicating a highly cooperative interaction involving multiple inhibitor sites. Sixteen other salts that were investigated also exhibited this pattern of inhibition. NaCl functions as a dead-end inhibitor without altering the number of MgATP hydrolyzed/electron transferred to substrate. The level of expressed inhibition is sensitive to MgATP concentration, the molar ratio of the MoFe-protein (Av1) to the Fe-protein (Av2), and total protein concentration. In addition, NaCl is an inhibitor of the MgATP-dependent, iron chelation of Av2. Although the inhibition is exhibited over the same salt concentration range as that for inhibition of substrate reduction, the pattern of inhibition is hyperbolic. A model based upon simple equilibrium interactions among the enzyme species, nucleotides, and inhibitor has been developed which quantitatively accounts for the observed effects of salt. In this model, the formation of the active complex between Av1 and Av2 is abolished by salts. Likewise, the apparent affinity of Av2 for MgATP is reduced. An additional prediction based upon the model is that the affinity between Av2 and Av1 is independent of nucleotide binding.
authors.affiliation: Department of Biochemistry, Michigan State University, East Lansing 48824.
authors.forename: T L
authors.initials: TL
authors.lastname: Deits
authors.forename: J B
authors.initials: JB
authors.lastname: Howard
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001395:Azotobacter / Q000201:enzymology*; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D009591:Nitrogenase / Q000037:antagonists & inhibitors*; D009994:Osmolar Concentration; D012492:Salts; D012965:Sodium Chloride / Q000494:pharmacology*
chemical_list: D012492:Salts; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D009591:Nitrogenase